Diagnosis of intoxications of piglets fed with Fusarium toxin-contaminated maize by the analysis of mycotoxin residues in serum, liquor and urine with LC-MS/MS

Concentrations of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites alpha-zearalenol (alpha-ZEL), beta-zearalenol (beta-ZEL), zearalanone (ZAN), alpha-zearalanol (alpha-ZAL), beta-zearalanol (beta-ZAL) and de-epoxy-deoxynivalenol (de-DON) in serum, liquor and urine of female piglets fed diets containing 0.01, 0.05, 0.08, 0.17 and 0.29mg ZEN/kg and 0.03, 0.59, 1.27, 2.01 and 4.52mg DON/kg during 29days of treatment were analysed. After 1, 3, 8, 15, 22 and 29days, four piglets per group were slaughtered. The simultaneous determination of all analytes was carried out using a sensitive and selective in-house-validated liquid chromatography tandem mass spectrometry (LC-MS/MS) method after sample preparation with Oasis (TM) HLB columns. ZEN, alpha-ZEL, DON and de-DON were detected in serum, whereas in liquor only ZEN, DON and de-DON were found at lower concentrations. In urine, all analytes were detected in considerably higher concentrations as in serum and liquor, whereby alpha- and beta-ZAL could only be detected sporadically. Apart from ZEN in liquor and alpha- and beta-ZAL in urine, the mycotoxin concentrations increased with increasing concentrations of Fusarium toxins in the diet. The toxin intake per kg body weight 3-4h prior to slaughtering correlated well with the DON and the sum of DON and de-DON concentrations in all three specimens as well as with the ZEN, alpha-ZEL and the sum of ZEN and metabolite concentrations in urine. Due to the high correlation between the dietary DON concentration and the DON (r=0.855) and the sum of DON and de-DON (r=0.870) concentration in serum, the exposure to DON can be evaluated. Moreover, serum levels of these toxins indicative of an exceeding of the guidance value in feed can be established using the corresponding regression equations. Strictly speaking, these relationships are only valid for the experimental conditions of the underlying experiment. For practical application of these relationships, the individual variation needs to be additionally considered. Effects of the duration of toxin exposure within the feeding groups were observed for ZEN, DON and de-DON in all specimens as well as for alpha-ZEL, beta-ZEL and ZAN in urine.